ANALYSIS AND ISOLATION OF SOLUBLE CD4 BINDING RETROVIRUS FROM THE PATIENTS WITH COLLAGEN DISEASES

本项目试图在外周血淋巴细胞和滑膜中分离具有HIV相关逆转录病毒CD4结合结构域的未知逆转录病毒。在RA患者的血清中，与HIV中gp120结合的可溶性sCD4的浓度明显高于正常人。逆转录酶聚合酶链反应（RT-PCR）方法扩增了类风湿性关节炎（RA）患者外周血淋巴细胞和鼻窦组织中HIV病毒gp120的非点突变区，无法检测到这种未知的逆转录病毒mRNA。采用荧光素差异显示法检测血清中sCD4浓度高的RA患者的疾病相关基因。本研究证实外周血淋巴细胞有分化带。对其中一条单链DNA进行亚克隆，并用自动DNA测序仪对其序列进行分析。DNA序列与人内源性逆转录病毒相似。目前，我们正通过定量RT-PCR进一步研究该基因在RA患者外周血淋巴细胞和滑膜中的表达情况。这可能是首次报道在RA患者外周血淋巴细胞中存在未知的人内源性逆转录病毒。进一步的研究将阐明这种内源性逆转录病毒与RA发病机制之间的关系。

This project try to isolate unknown retrovirus which had CD4 binding domein of HIV related retrovirus in the peripheral blood lymphocytes and synovium. The concentration of to soluble (sCD4) which bound to gp120 in HIV,was significant higher in sera from patients with RA than normal individuals. This unknown retrovirus mRNA was not able to detect by reverse transcriptase polymerase chain reaction (RT-PCR) method using primers which amplified the non point mutated region of gp120 in HIV in the peripheral blood lymphocytes and sinovial tissue from the patients with rheumatoid arthritis (RA). Furthemore we employed Fluorescein Differencial Display method to detect the disease associated genes in the patients with RA with high concentration of sCD4 in sera. This study demonstrated the some differentiated bands in the periferal blood lymphocytes. One of these single stranded DNA bands was subcloned and analyzed the DNA sequence by the automatic DNA sequencer. The DNA sequence was similar to an human endogenous retrovirus. Now we are further studing the expression of this gnen in the peripheral lymphocytes and synovium from the patientes with RA by quantitative RT-PCR.This might be a first report that demonstrated the existence of unknown human endogenous retrovirus in periferal blood lymphocytes from the patients with RA.Further investigation will clarify the association between this endogenous retrovirus and pathogenesis of RA.